Regulatory circuitry of RNA polymerase II c-terminal domain phosphatase Fcp1 in Saccharomyces cerevisiae

在细胞中，生命所必需的信息以称为基因的单位存储在DNA上。每个基因都包含生产一种蛋白质所需的信息，其中一些蛋白质在任何时候都是必需的，而另一些蛋白质只在某些条件下才需要。因此，蛋白质生产的控制是维持正常细胞功能的关键过程。 细胞中所有蛋白质的产生都始于一个称为转录的过程，在此过程中，所需基因的拷贝被制作出来。这是一个重要的过程，因为如果使用DNA模板本身，重要信息的损坏和丢失的机会将大大增加。RNA聚合酶II（RNAPII）是负责大多数基因转录的酶。它与其他RNA聚合酶不同，因为它含有一个称为C-末端结构域（CTD）的尾部区域。CTD的功能就像一个港口，在那里与RNAPII一起工作的蛋白质在转录的不同阶段根据需要停靠。CTD必须确保在任何给定时间只有正确的蛋白质结合，因为过早或延迟的结合会导致不稳定或无功能的DNA拷贝。CTD通过改变其形状来实现这一点，仅允许正确形状的蛋白质在给定时间内适合。Fcp1是一种在转录的各个阶段帮助CTD重塑过程的蛋白质。虽然Fcp1的作用是已知的，但Fcp1如何识别何时重塑CTD仍然是未知的，这将是本研究提案的重点。 我们已经进行了遗传筛选，重点是已知在转录调控中发挥作用的基因，以确定可能的Fcp1调节剂。我们已经生成了一个潜在候选者的列表，通过观察抑制这些候选者的功能如何导致CTD重塑缺陷和一般转录缺陷来进行后续研究。我们还将确定它们对Fcp1的调控是否仅影响特定基因或所有基因。如果只有特定的基因受到影响，这将表明有多种蛋白质调节Fcp1的功能，这就要求我们识别它们，以更好地了解Fcp1如何调节转录过程。

In the cell, information essential for life is stored on the DNA in units called genes. Each gene contains the information required for the production of one protein, some of which are required at all times, while others only under certain conditions. As a result, control of protein production is a critical process for maintenance of normal cellular function. All protein production in the cell starts with a process called transcription during which a copy of the required gene is made. This is an important process because if the DNA template itself were to be used, the chances of damage and loss of vital information are greatly increased. RNA Polymerase II (RNAPII) is the enzyme responsible for the transcription of the majority of genes. It is different from other RNA polymerases as it contains a tail region called the C-terminal domain (CTD). The CTD functions like a port where proteins that work with RNAPII dock as required at different stages of transcription. The CTD must ensure that only the correct proteins bind at any given time as premature or delayed binding leads to unstable or non-functional DNA copies. The CTD achieves this is by changing its shape, allowing only correctly shaped proteins to fit at a given time. Fcp1 is a protein that aids in the CTD remodeling process at various stages of transcription. While the role of Fcp1 is known, how Fcp1 recognizes when to remodel the CTD remains unknown and will be the focus of this research proposal. We have performed a genetic screen focusing on genes already known to play a role in transcription regulation to identify possible Fcp1 regulators. We have generated a list of potential candidates to follow up on by looking at how inhibiting the function of these candidates leads to CTD remodeling defects and general transcription defects. We will also determine if their regulation of Fcp1 affects only specific genes or all genes. If only specific genes are affected, it will suggest that multiple proteins regulate Fcp1 function, requiring that we identify them to better understand how the transcription process is regulated by Fcp1.